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Laser Capture Microdissection Protocol for Xylem Tissues of Woody Plants

2017-01-04Journal article Open access
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dc.contributor.authorBlokhina, Olga
dc.contributor.authorValerio, Concetta
dc.contributor.authorSokołowska, Katarzyna
dc.contributor.authorZhao, Lei
dc.contributor.authorKärkönen, Anna
dc.contributor.authorNiittylä, Totte
dc.contributor.authorFagerstedt, Kurt
dc.date.accessioned2017-01-23T15:24:10Z
dc.date.available2017-01-23T15:24:10Z
dc.date.issued2017-01-04
dc.description.abstractLaser capture microdissection (LCM) enables precise dissection and collection of individual cell types from complex tissues. When applied to plant cells, and especially to woody tissues, LCM requires extensive optimization to overcome such factors as rigid cell walls, large central vacuoles, intercellular spaces, and technical issues with thickness and flatness of the sections. Here we present an optimized protocol for the laser-assisted microdissection of developing xylem from mature trees: a gymnosperm (Norway spruce, Picea abies) and an angiosperm (aspen, Populus tremula) tree. Different cell types of spruce and aspen wood (i.e., ray cells, tracheary elements, and fibers) were successfully microdissected from tangential, cross and radial cryosections of the current year's growth ring. Two approaches were applied to achieve satisfactory flatness and anatomical integrity of the spruce and aspen specimens. The commonly used membrane slides were ineffective as a mounting surface for the wood cryosections. Instead, in the present protocol we use glass slides, and introduce a glass slide sandwich assembly for the preparation of aspen sections. To ascertain that not only the anatomical integrity of the plant tissue, but also the molecular features were not compromised during the whole LCM procedure, good quality total RNA could be extracted from the microdissected cells. This showed the efficiency of the protocol and established that our methodology can be integrated in transcriptome analyses to elucidate cell-specific molecular events regulating wood formation in trees.pt_PT
dc.description.sponsorshipAcademy of Finland grants: (#251390, #283245); Bio4Energy (Swedish Programme for Renewable Energy); Kempe Foundation fellowship.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationBlokhina O, Valerio C, Sokołowska K, Zhao L, Kärkönen A, Niittylä T and Fagerstedt K (2017) Laser Capture Microdissection Protocol for Xylem Tissues of Woody Plants. Front. Plant Sci. 7:1965. doi: 10.3389/fpls.2016.01965pt_PT
dc.identifier.doi10.3389/fpls.2016.01965pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.7/729
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherFrontiers Mediapt_PT
dc.relation.publisherversionhttp://journal.frontiersin.org/article/10.3389/fpls.2016.01965/fullpt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectcryosectionpt_PT
dc.subjectlaser capture microdissectionpt_PT
dc.subjectray cellspt_PT
dc.subjectRNA integritypt_PT
dc.subjecttracheidspt_PT
dc.subjectxylem fiberspt_PT
dc.titleLaser Capture Microdissection Protocol for Xylem Tissues of Woody Plantspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage14pt_PT
oaire.citation.startPage1pt_PT
oaire.citation.titleFrontiers in Plant Sciencept_PT
oaire.citation.volume7pt_PT
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT

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